Presence of anti-Sm reactivity in autoimmune mouse strains

The investigation of the fine specificities of antinuclear antibodies (ANAs) has been fruitful in terms of the nosology and immunopathogenesis of human autoimmune syndromes. Particular reactivities serve as “markers,” in that patients with certain syndromes have a much higher incidence of such ANAs than do patients with other diseases. In this category is the almost exclusive against the nuclear acidic protein Sm. Reactivity to Sm can be detected by precipitation in agar, complement fixation, or passive hemagglutination (1,2). Autoimmune mouse strains have also provided a fertile field for the investigation of the basic phenomena of self-activity. In particular, the NZB strain and its hybrid NZB x NZW have been considered excellent models for human SLE and have therefore been studied in great detail (3,4). In addition, Murphy et al at The Jackson Laboratory, Bar Harbor, Maine, have developed several new inbred mouse strains that spontaneously develop SLE-like syndromes (5,6). These are the BXSB strain, which has a male dominant disease characterized by little antiative DNA antibody; the MRL/1, which develops massive, nonmalignant lymphadenopathy, associated with enormous increases in serum immunoglobulin levels and fulminant renal disease; and the MRL/n, which does not develop SLE-like disease until well into the 2nd yr of life, but like the MRL/1 develops high titers of ANA and fatal glomerulonephritis. The MRL/1 differs from MRL/n in only about 10 percent of its genome, including the gene responsible for the MRL/1’s lymphoproliferation. In the current study, we have used the technique of double immunodiffusion (ID) in agarose with standard human reference sera (of known ANA specificity) to survey a large number of mice from the NZB x NZW, MRL/1, MRL/n, BXSB, and other strains. We report here the finding of the anti-Sm marker” antibody almost uniquely in MRL/1 and MRL/n animals. These two related strains may serve as experimental models to explore the mechanism stimulating the production of this unique autoantibody in SLE.     

Compositional analysis of the collagenous bone matrix. A study on adult normal and osteopenic bone tissue

The collagenous constituents of mature bone of 30 individuals 22-93 years of age were studied by post-mortem morphological and biochemical analysis. Morphometric evaluation of the second lumbar vertebral body revealed striking interindividual differences in bone mass, mean trabecular density and mean trabecular thickness. Collagen extracted from vertebral trabecular bone by limited pepsin digestion consisted mainly of collagen I (92%) and collagen V (8%). Immunohistochemistry revealed a distinct distribution of these two collagen types within the bone matrix. The degree of lysyl hydroxylation of the alpha 2(I) collagen chain correlated inversely with the trabecular bone volume (TBV) and with the mean trabecular plate density. This correlation was statistically significant for the entire study group as well as for the female and male subgroups. Within the female subgroup, the lysyl hydroxylation/TBV ratio was higher in postmenopausal than in premenopausal women and was highest in women with established osteoporosis. No significant correlation was found between the level of lysyl hydroxylation and the age of the patients. The alpha 1(I) collagen chain showed a nearly constant degree of lysyl hydroxylation in all 30 samples. The results provide convincing evidence that morphometric changes associated with osteopenia in adult bone are accompanied by an altered level of lysyl hydroxylation of the alpha 2(I)-chain of collagen I. The biochemical alterations observed may be responsible for the deposition of a deficient bone matrix in osteopenic conditions.     

Carcinogenesis in tissue culture. 28. Comparison of various effects of a chemical carcinogen, 4-nitroquinoline I-oxide, on normal human cells and on normal mouse cells in culture.

A comparison of the effects on cultured cells of a chemical carcinogen, 4-nitroquinoline 1-oxide (4NQO), between normal human cells which were resistant to malignant transformation with 4NQO and normal mouse cells rather easily transformable was carried out. We studied the following effects of 4NQO on normal human and normal rodent cells; 1) cytotoxicity, 2) DNA, RNA and protein synthesis, 3) incorporation of 4NQO into cells and time course changes of the drug bound with macromolecular substances in cells, 4) DNA repair synthesis, and 5) chromosomal changes. Our results demonstrated that there were no differences in cytotoxicity and inhibition of cellular macromolecular syntheses between human and mouse cells. On the other hand, significant differences were noted in DNA repair synthesis and chromosomal aberrations between human and rodent cells. These differences suggest that mouse cells are easily transformed into neoplastic cells with chemical carcinogens as compared with human cells.     

X射线影像在正常骨组织及骨疾病诊断中的应用分析

背景:X射线影像诊断已经成为临床影像诊断中重要的组成部分,是骨关节系统临床诊治最常用的方法。目的:对X射线影像在正常骨组织及骨疾病诊断中的应用研究文献进行多层次探讨分析。方法:以电子检索的方式对CNKI数据库2002/2011X射线影像在正常骨组织及骨疾病诊断中的应用研究文献进行检索分析,采用检索词为骨组织;髋关节;膝关节;肩关节;肘关节;X射线影像。骨关节系统是人体的承重系统及运动系统,结构复杂多样化,因此骨组织X射线影像表现也呈现复杂多样性,骨组织X射线影像诊断为临床诊断治疗提供重要的辅助依据。结果与结论:人体组织结构存在密度和厚度差异的不同,X射线穿透人体时被吸收的量也不同,以致剩余的X射线量也不同,剩余的X射线经过成像介质的显像或计算机的重建过程,在X射线片上就能显示具有黑白对比、层次差异的X射线图像。X射线影像不仅应用于正常骨组织的研究,而且应用于骨组织的疾病诊断。     

Presence in mouse bone marrow of mediator-dependent cytotoxic effector cells against tumors

The role of bone marrow cells as cytotoxic effector cells against tumor cells was examined in vitro. Bone marrow cells from normal C3H/He mice could lyse murine MM46 tumor cells in the presence of wheat germ agglutinin or syngeneic antitumor antibody. Bone marrow cells alone did not cause lysis. Eleven other lectins (concanavalin A, phytohemagglutinin, pokeweek mitogen, Lens culinaris hemagglutinin, Ulex europaeus agglutinin, peanut agglutinin, Bandeiraea simplicifolia agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin, Limulus polyphemus agglutinin and Helix aspersa agglutinin) did not induce cytolysis. Enrichment of polymorphonuclear leukocytes (PMNs) by the method of free flow electrophoresis enhanced the activity of WGA-dependent cytolysis. These results suggest that PMNs series in the bone marrow of normal mice can lyse tumor cells with appropriate mediators.     

Nonrandom inclusion of H-2K and H-2D antigens in Friend virus particles from mice of various strains

Friend murine leukemia virus (FV), isolated from infectious serum of several mouse strains, has been examined for the presence of H-2 antigens. Following banding of the virus on a discontinuous sucrose gradient, pelleting, and disruption with Nonidet P-40 detergent, virus preparations were tested for their capacity to inhibit the lysis of target cells mediated by various anti-H-2K or anti-H-2D antisera. Virus from mice homozygous for the H-2b, H-2d, H-2g H-2k, and H-2ol haplotypes or heterozygous for the H-2g/H-2k, H-2b/H-2d, and H-2b/H-2k haplotypes was used. Of the six H-2D or H-2K alleles examined, the products of only two, H-2Db and H-2Kk were detected. Virus preparations contained no, one, or both antigens, depending on the genotype of the host animal. Control preparations from normal mouse serum and preparations in which the virus had not been disrupted demonstrated no H-2 activity. Furthermore, attempts to neutralize FV spleen focus forming activity with anti-H-2Db or anti-H-2Kk antisera yielded negative results.     

The degree of mineralization in normal bone tissue estimated as the phosphorus to hydroxyproline ratio.

The degree of mineralization can be evaluated from the phosphorus to hydroxyproline ratio in bone biopsies. A small but significant rise in the ratio with increasing age was observed in 137 persons without bone diseases, aged from 20 to 89 years. No significant difference between men and women was found with regard to the phosphorus to hydroxyproline ratio. The study indicates that with increasing age, in addition to senile osteopenia, there is also a change in the form of an increase in the degree of mineralization.     

The normal levels of fluorine in the bone tissue of Japanese subjects

The variation of fluorine concentration by sex and age in bones of Japanese subjects was estimated. The subjects had lived in districts virtually with no fluoride in drinking water and air. The rib bone and the iliac crest were selected for investigation. The fluorine concentration in bones was about 100 ppm in dry (fat-free) weight and 200-230 ppm in ash weight from birth to 19 in both sexes, and increased in their twenties, reaching a plateau level in the 60's. The fluorine concentration in rib bones, however, was 610 ppm in dry (fat-free) weight and 1100 ppm in ash weight, which was slightly higher than that in ilia with 530 ppm in dry (fat-free) weight and 960 ppm in ash weight, respectively. The bones of males contained more fluorine than those of females. The concentration of Ca, P and Mg in bone tissue did not correlate with fluorine concentration.     

Incidence and specificities of IgA and IgM anti-AgG autoantibodies in various mouse strains and colonies

Mice, greater than 20 wk old, were tested for the presence of anti-IgG autoantibodies by agglutination and radioimmunoassay. IgA and IgM anti- IgG were found in the 129/Sv, C57BL/6, and DBA/2 strains from the local colony at the International Institute of Cellular and Molecular Pathology (ICP), at the Institut Pasteur de Paris (IP), and in the endotoxin-resistant C3H/He strain of ICP. These strains were negative at Iffa Credo (IC), and at The Jackson Laboratory (JL). Among 33 strains from the latter colony, 129/J, AKR/J, CBA/J, C57L/J, and NZB/BinJ were positive. All were specific pathogen-free and, excepting the NZB/BinJ, are not known to develop systemic autoimmune disorders. These differences between colonies suggest an influence of the environment on the production of anti-IgG. Evidence for the role of an infectious agent was provided by the fact that germ-free DBA/2 were negative in contrast to their SPF relatives. Strains which were positive at ICP and IP for anti-IgG had four-times higher serum levels of total IgA and two-times higher levels of total IgG than the corresponding negative strains from IC and JL. The anti-IgG titers differed markedly from one strain to the other in the same environment; e.g., in mice from ICP, BALB/c mice produced 40-times less anti-IgG than 129/Sv. IgA anti-IgG occurred only in high producers of anti-IgG. In these animals, the proportion of IgA vs. IgM anti-IgG was very different from one group to the other; C57BL/6 had mainly IgM anti-IgG, DBA/2 mainly IgA anti-IgG, and 129/Sv both IgM and IgA anti-IgG. The IgA anti-IgG from 129/Sv, 129/J, NZB/BinJ, C57L/J, DBA/2, and C3H/He had restricted hetero-, iso-, and allotypic specificities. It reacted only with mouse IgGa2, but not with the Ig-1b allotype. C57BL/6 also had IgA anti-IgG with a narrow specificity, but directed against IgG1 without allotypic restriction. In contrast to the specificity of IgA anti-IgG, the antibody activity of IgM anti-IgG was much broader, except in the 129/Sv and 129/J strains where IgM anti-IgG shared the same narrow specificity with IgA.     

Inhibition of growth of mouse ehrlich ascites by normal tissue extracts

A dialyzable component from the aqueous extracts of mouse skeletal muscle and liver inhibited in vitro growth of a mouse Ehrlich ascites. A similar component was not detected in extracts of spleen, kidney, lung, skin, serum or small intestine. The muscle component appeared to be different from that of the liver in its resistance to heat and its stability in the culture medium. Both components however were stable on storage at 4, 23 and 37 degrees C for 72 h. Intraperitoneal injection of the muscle and liver component into mice previously innoculated with Ehrlich ascites significantly decreased tumor incidence in these animals as compared with the control.     

Presence of serum and tissue forms of N-acetyl-beta-glucosaminidase in urine from patients with renal disease.

1. The A, B, I1 and I2 forms of N-acetyl-beta-glucosaminidase present in urine, serum, kidney, liver and cerebral spinal fluid were separated on DEAE-cellulose and their presence confirmed by cellogel electrophoresis. The relative activities of each enzyme were determined by integrating the area under the elution peaks. 2. Serum A-form was eluted at a lower molarity of chloride than liver A-form and this was designated the As-form to distinguish it from the A-form of N-acetyl-beta-glucosaminidase found in liver and kidney. 3. The P-form of N-acetyl-beta-glucosaminidase present in the serum of a group of pregnant women was not detectable in urine samples from the same women. 4. Urinary NAG activities were found to be abnormally high in patients with impaired renal function. 5. The activity of both N-acetyl-beta-glucosaminidases A and B increased in pathological urines. The higher the total N-acetyl-beta-glucosaminidase activity excreted the higher the % of activity of the B-form present. 6. In a number of patients with haematuria an A-form similar to the serum As-form was present in the urine.     

Resorption of implanted bone prepared from normal and warfarin-treated rats.

Bone that was virtually depleted of the vitamin K-dependent protein, osteocalcin, and 93% reduced in the concentration of its characteristic amino acid, gamma-carboxyglutamic acid, was obtained from rats treated with warfarin for 6 wk. Osteocalcin-deficient bone particles were resistant to resorption when implanted subcutaneously in normal rats. The relative resorption was 60% of control bone, as measured by histomorphometry as percent of bone particles in the field. Additionally, the number of multinucleated cells around the bone particles was reduced by 54%. These data suggest that osteocalcin is an essential component for bone matrix to elicit progenitor-cell recruitment and differentiation necessary for bone resorption.     

Microcalorimetric studies on metabolism of hepatic tissue I. A methodological study of normal tissue

In the present study, the heat production of liver biopsies (5-8 mg) was measured by a microcalorimetric technique. Tissue incubated in Leibowitz L-15 medium (L-15) showed a higher metabolic rate compared to tissue incubated in a medium without substrate 2.8 microW/mg and 1.75 microW/mg, respectively. Heat production was found to be related to weight density. No difference in the metabolic rate was found after organ perfusion in comparison to nonperfused liver. Storage in medium L-15 at 4 degrees C caused a lower rate of heat production, but if the tissue was stored in an electrolyte balance solution without substrate, no difference was seen compared to fresh tissue. Recording heat production with the present calorimetric technique is relatively simple and rapid and allows measurement of small samples.     

Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow

We have resolved B220+ IgM- B-lineage cells in mouse bone marrow into four fractions based on differential cell surface expression of determinants recognized by S7 (leukosialin, CD43), BP-1, and 30F1 (heat stable antigen). Functional differences among these fractions can be correlated with Ig gene rearrangement status. The largest fraction, lacking S7, consists of pre-B cells whereas the others, expressing S7, include B lineage cells before pre-B. These S7+ fractions, provisionally termed Fr. A, Fr. B, and Fr. C, can differentiate in a stromal layer culture system. Phenotypic alteration during such culture suggests an ordering of these stages from Fr. A to Fr. B to Fr. C and thence to S7- pre-B cells. Using polymerase chain reaction amplification with pairs of oligonucleotide primers for regions 5' of JH1, DFL16.1, and Jk1, we find that the Ig genes of Fr. A are in germline configuration, whereas Fr. B and C are pro-B cell stages with increasing D-J rearrangement, but no V-D-J. Finally, functional analysis demonstrates that the proliferative response to IL-7, an early B lineage growth factor, is restricted to S7+ stages and, furthermore, that an additional, cell contact-mediated signal is essential for survival of Fr. A.     

Flow cytometry of needle aspirates from bone and soft tissue tumors.

Fine needle aspiration specimens from 29 patients with bone and soft tissue neoplasms were analyzed by flow cytometry for DNA index and cell cycle analysis to determine whether such studies were helpful in cytologic diagnosis. Of 15 cases initially cytologically diagnosed as benign, 14 had a DNA index of 1.0, indicating a diploid population. The remaining case diagnosed as cytologically benign had a DNA index of 1.3. Further tissue from this tumor revealed an osteogenic sarcoma. Of the 14 cases initially diagnosed as malignant, 12 were hyperdiploid. Cell cycle analysis showed that malignant tumors had a higher proportion of cells in S phase (15.2% +/- 8.7%) than benign tumors (6.9% +/- 1.6%). Furthermore, high-grade malignancies had a significantly greater number of cells in S phase (18.5% +/- 1.5%) than low-grade tumors (9.9% +/- 6.3%).     

Reduction of soft tissue deposition in normal triplets.

In a previous study, birth weight projections obtained with Rossavik growth models were systematically greater than actual birth weights in triplets but not in singletons. To investigate the cause of this overestimation, average mean percent deviations for head circumference (HC), abdominal circumference (AC), thigh circumference (ThC), and weight (WT) at different times in the third trimester were studied in 13 normal triplet and 20 normal singleton fetuses using individual growth curve standards. Average mean percent deviation values for HC in both singletons and triplets were close to zero throughout the third trimester. Average mean percent deviation values for AC in both singletons and triplets were similar, and remained relatively constant. Average mean percent deviation values for ThC in triplets decreased toward the end of the third trimester, but those for singletons increased. Average mean percent deviation values for estimated weight did not change during the third trimester in singletons whereas those in triplets decreased. The difference in average mean percent deviation values for estimated weight between singletons and triplets in the 32- to 36-week interval was statistically significant. There were no significant differences between actual and predicted measurements for HC in both singletons and triplets at birth. However, actual birth measurements were systematically less than predicted birth measurements for AC and ThC in singletons, and for AC, ThC, and WT for triplets. These systematic differences were significantly larger in triplets for ThC and WT. Nutrition Score values, direct measures of subcutaneous tissue, were significantly lower in triplets compared to singletons.(ABSTRACT TRUNCATED AT 250 WORDS)